원문정보
초록
영어
A cDNA encoding manganese peroxidase isozyme H4 (MnPH4), isolated from Phanerochaete chrysosporium, was expressed in Pichia pastoris, under the control of the alcohol oxidase I promoter. The recombinant MnPH4 is efficiently secreted into the medium upon hemin supplementation, at a maximum concentration of 500 U/L, and the purified rMnPH4 was used to decolorize the triphenylmethane dye, Malachite green (MG). Response surface methodology (RSM) was employed to optimize different operational parameters on the decolorization of MG. RSM showed that the optimized variables viz. enzyme (0.662 U), MnSO4 (448 mM) and hydrogen peroxide (159mM) decolorized 100 ppm of MG completely in 3 hours. UV-VIS spectra, HPLC, GC-MS and LC-ESI MS analysis showed the degradation of MG. Finally, treatment of MG with rMnPH4 resulted in the complete removal of toxicity which was checked under in vitro conditions.